Please wait a minute...
Journal of Integrative Agriculture  2026, Vol. 25 Issue (2): 721-733    DOI: 10.1016/j.jia.2025.02.042
Horticulture Advanced Online Publication | Current Issue | Archive | Adv Search |
An integrate methods to improve the high efficiency of embryo rescue breeding in seedless grapes

Xi Chen1*, Khalid Ayesha1*, Xue Wen1, Yanan Zhang1, Mengru Dou1, Kexuan Jia1, Yong Wang2, Yuling Li2, Feng Sun2, Guotian Liu1#, Yan Xu1#

1 State Key Laboratory of Crop Stress Resistance and High-Efficiency Production/Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China, Ministry of Agriculture and Rural Affairs/College of Horticulture, Northwest A&F University, Yangling 712100, China

2 Xinjiang Uygur Autonomous Region Research and Development Center for Facility Agriculture and Specialty Agriculture, Shanshan 838200, China

Highlights

Optimization of culture media for embryo development and germination in seedless grapes demonstrated that supplementation with zeatin and auxin significantly enhances both embryo development and germination rates.

Refinement of the conversion medium for abnormal plantlets in embryo rescue, combined with establishment of a regeneration system for inducing adventitious buds from these plantlets, enabled the recovery of 35% of otherwise abnormal plantlets, thereby improving seedling formation efficiency in embryo rescue.

Download:  PDF in ScienceDirect  
Export:  BibTeX | EndNote (RIS)      
摘要  
胚挽救技术在培育无核葡萄新品种的过程中扮演着至关重要的角色。为了提高无核葡萄胚挽救育种效率,本研究配置了22个杂交组合,研究不同亲本基因型、植物激素在胚发育和萌发培养基中对胚挽救效率的影响,并对畸形苗的挽救进行研究。结果表明:‘红宝石无核’、‘底莱特’、‘火州黑玉’、 ‘紫甜无核’和‘郑艳无核’ 适合作母本材料;‘紫甜无核’、‘沈农香丰’、‘红旗特早’和‘瑰宝’ 适合作父本材料;其中杂交组合‘红宝石无核×沈农香丰’ 和 ‘红宝石无核×紫甜无核’的胚挽救效率最高,胚发育率分别为  55.05% and 59.76%,并分别获得了 1348 株和 2235 株杂种后代;在MM3 + 0.2 mg・L⁻¹ IAA(吲哚 - 3 - 乙酸)胚发育培养基中添加 1.0 mg・L⁻¹ ZT(玉米素)时,‘红宝石无核×紫甜无核’胚的发育率最高达 64.73%;在胚萌发WPM培养基中,添加 0.2 mg・L⁻¹ ZT + 0.2 mg・L⁻¹ IAA 使杂交组合‘火州黑玉×阳光玫瑰’的萌发率达到最高,为 85.71%;同时,通过直接转化法和胚轴诱导不定芽再生途径,将3365株胚挽救畸形苗成功转化为正常苗1234株。最后成功移栽了4287株杂种植株。本研究为提高无核葡萄胚挽救育种效率提供了理论依据,为未来的无核葡萄育种计划提供了宝贵的材料。




Abstract  
The embryo rescue technique plays an essential role in developing new seedless grape varieties.  To enhance the efficiency of seedless grape embryo rescue breeding, this study evaluated 22 hybrid combinations and systematically investigated the effects of parental genotypes and plant hormones on embryo development and germination.  Additionally, an in-depth analysis was conducted on the conversion of abnormal plantlets.  Results indicate that ‘Ruby Seedless’, ‘Delight’, ‘Huozhouheiyu’, ‘Zitian Seedless’, and ‘Zhengyan Seedless’ are suitable as maternal parents, whereas ‘Zitian Seedless’, ‘Shennongxiangfeng’, ‘Hongqitezao’, and ‘Guibao’ perform optimally as paternal parents.  Among these, the crosses ‘Ruby Seedless×Shennongxiangfeng’ and ‘Ruby Seedless×Zitian Seedless’ exhibited the highest embryo rescue efficiency, with embryo development rates of 55.05 and 59.76%, yielding 1,348 and 2,235 viable plantlets, respectively.  When 1.0 mg L–1 zeatin (ZT) was added to the MM3 medium supplemented with 0.2 mg L–1 indole-3-acetic acid (IAA), the embryo development rate of ‘Ruby Seedless×Zitian Seedless’ increased by 64.73%.  In the WPM germination medium, supplementation with 0.2 mg L–1 ZT and 0.2 mg L–1 IAA resulted in the highest germination rate of 85.71% for the hybrid combination ‘Huozhouheiyu×Shine Muscat’.  Furthermore, 3,365 abnormal plantlets were rescued via direct transformation and hypocotyl-induced adventitious bud regeneration, among which 1,234 were transformed into normal plantlets.  Following hybridization, a total of 4,287 plants were successfully acclimatized and transplanted.  This study provides theoretical insights to improve the efficiency of embryo rescue breeding in seedless grapes and offers valuable genetic resources for future breeding programs.
Keywords:  seedless grape       embryo rescue        parental genotype        plant hormones        abnormal plantlets  
Received: 07 November 2024   Accepted: 26 January 2025 Online: 20 February 2025  
Fund: 

This work was supported by the China Agriculture Research System of MOF and MARA (CARS-29-yc-3).

About author:  Xi Chen, E-mail: xichen@nwafu.edu.cn; Khalid Ayesha, E-mail: aleenakhalidk25@gmail.com; #Correspondence Yan Xu, E-mail: yan.xu@nwafu.edu.cn; Guotian Liu, E-mail: gtliu@nwafu.edu.cn *These authors contributed equally to this study.

Cite this article: 

Xi Chen, Khalid Ayesha, Xue Wen, Yanan Zhang, Mengru Dou, Kexuan Jia, Yong Wang, Yuling Li, Feng Sun, Guotian Liu, Yan Xu. 2026. An integrate methods to improve the high efficiency of embryo rescue breeding in seedless grapes. Journal of Integrative Agriculture, 25(2): 721-733.

Benke A P, Krishna R, Samarth R R, Dhumal S S, Ansari W A, Shelke P V, Dukare S S, Singh M. 2021. Development of an embryo germination protocol for shy-seeded grape (Vitis vinifera L.). Plant Genetic Resources Characterization and Utilization19, 252–260.

Bouquet A, Davis H P, Danglot Y, Rennes C. 1989. In ovulo and in vitro embryo culture for breeding seedless table grapes (Vitis vinifera L.). Agronomie9, 565–574.

Bruce P B, James N M. 1994. Promoting stenospermic grape seed trace development and germination with plant growth regulators. Journal of the American Society for Horticultural Science, 119, 719–716.

Cain D W, Emershad R L, Tarail R E. 1983. In-ovulo embryo culture and seedling development of seeded and seedless grapes (Vitis vinifera L.). Vitis22, 9–14.

Celik H, Ilbay A K. 2003. The use of in ovulo embryo culture for cross-breeding studies of empty-Seeded table grape cultivars. Acta Horticulturae603, 189–193.

Celso Valdevino P, David W R, Richard L E. 1995. Influence of grape genotype, ripening season, seed trace size, and culture date on in ovule embryo development and plant formation. Bragantia54, 237–249.

Chiaromonte E, Bottalico G, Lanotte P, Campanale A, Montilon V, Morano M, Saponari A, Pirolo C S, Gerin D, Faretra F, Pollastro S, Savino V N. 2023. A large-scale validation of an improved embryo-rescue protocol for the obtainment of new table-grape seedless genotypes. Plants12, 3469.

Chu Y N, Li M, Li R N, Zhang K Z, Qiu P P, Yuan X J, Han Y L, Liu X Y, Xu Y, Liu G T. 2023. Embryo rescue breeding of new cold-resistant, seedless grapes. Horticulturae, 9, 992.

Cui X Y, Liu Q Y, Luo Y J, Zhu P P, Guan P Y, Zhang J X. 2024. Study on influencing factors of embryo rescue and germplasm innovation in seedless grape. Plant CellTissue and Organ Culture157, 24.

David W R, Richard L E, Ronald E T. 2000. A stenospermocarpic, seedless vitis vinifera×vitis rotundifolia hybrid developed by embryo rescue. Hortscience35, 732–734.

Ding H F, Qi G M. 2001. Ovules culture and plant formation of hybrid progeny of seedless grape. Journal of Agricultural in the Tropics and Subtropics102, 147–152.

Emershad R L, Ramming D W. 1984. In-ovulo embryo culture of Vitis vinifera L. C.V. ‘Thompson Seedless’. American Journal of Botany71, 873–877.

Giancaspro A, Mazzeo A, Carlomagno A, Gadaleta A, Somma S, Ferrara G. 2022. Optimization of an in vitro embryo rescue protocol for breeding seedless table grapes (Vitis vinifera L.) in Italy. Horticulturae8, 121.

Goldy R G, Amborn U. 1987. In vitro culturability of ovules from 10 seedless grape clones. Hortscience22, 952.

Gray D J, Fisher L C, Mortensen J A. 1987. Comparison of methodologies for in ovulo embryo rescue of seedless grapes. Hortscience22, 1334–1335.

Gray D J, Mortensen J A, Benton C, Durham R E, Moore G A. 1990. Ovule culture to obtain progeny from hybrid seedless bunch grapes. Journal of the American Society for Horticultural Science115, 1019–1024.

Guo Y S, Zhao Y H, Li K, Liu Z D, Lin H, Guo X W, Li C X. 2011. Embryo rescue of crosses between diploid and tetraploid grape cultivars and production of triploid plants. African Journal of Biotechnology10, 19005–19010.

Hiroyuki Y, Tadayuki H, Hisashi S. 1995. In vitro culture of embryos obtained by crossing tetraploid grape cultivar ‘Kyoho’ with diploid cultivars. Journal of the Japanese Society for Horticultural Science63, 719–724.

Ji W, Li Z Q, Yao W K, Gong P J, Wang Y J. 2013a. Abnormal seedlings emerged during embryo rescue and its remedy for seedless grape breeding. Korean Journal of Horticultural Science & Technology31, 483–489.

Ji W, Li Z Q, Zhou Q, Yao W K, Wang Y J. 2013b. Breeding new seedless grape by means of in vitro embryo rescue. Genetics and Molecular Research12, 859–869.

Ji W, Wang Y J. 2013. Breeding for seedless grapes using Chinese wild Vitis spp. II. In vitro embryo rescue and plant development. Journal of the Science of Food and Agriculture93, 3870–3875.

Jiao Y T, Li Z Q, Xu K Y, Guo Y R, Zhang C, Li T M, Jiang Y C, Liu G T, Xu Y. 2018. Study on improving plantlet development and embryo germination rates in in vitro embryo rescue of seedless grapevine. New Zealand Journal of Crop and Horticultural Science46, 39–53.

Li G R, Ji W, Wang G, Zhang J X, Wang Y J. 2014. An improved embryo-rescue protocol for hybrid progeny from seedless Vitis vinifera grapes×wild Chinese Vitis species. In Vitro Cellular & Developmental Biology (Plant), 50, 110–120.

Li G R, Wang G, Wang Y J. 2013. Embryo rescue to obtain progeny from seedless grapes (Vitis vinifera L.)×wild Chinese Vitis species. Journal of Food Agriculture and Environment, 11, 1058–1062.

Li S S, Liu K K, Yu S S, Jia S S, Chen S, Fu Y H, Sun F, Luo Q W, Wang Y J. 2020. The process of embryo abortion of stenospermocarpic grape and it develops into plantlet in vitro using embryo rescue. Plant CellTissue and Organ Culture143, 389–409.

Liu Q, Zhang J X, Wang Y J, Yu D, Xia H. 2016. Breeding for cold-resistant, seedless grapes from Chinese wild Vitis amurensis using embryo rescue. New Zealand Journal of Crop and Horticultural Science44, 136–151.

Liu S M, Sykes S R, Clingeleffer P R. 2003. Improved in ovulo embryo culture for stenospermocarpic grapes (Vitis vinifera L.). Australian Journal of Agricultural Research54, 869–876.

Loomis N H, Weinberger J H. 1979. Inheritance studies of seedlessness in grapes. Journal of the American Society for Horticultural Science104, 181–184.

Luo Y J, Cui X Y, Zhu P P, Zhang J X. 2023. Effects of parental genotypes on embryo rescue of seedless grape hybrid. New Zealand Journal of Crop and Horticultural Science9, 1–12.

Mazdak R, Marandi R J, Baneh H D, Hosseini B, Behnam H, Reza D. 2013. Effect of paternal genotypes sprays with BA and IAA concentration on embryo rescue of F1 progenies from ‘Askari’ (Vitis vinifera L.) cultivar. Journal of Agricultural Science and Technology15, 1023–1032.

Ponce M T, María T P, Agüero C B, Gregori M T, Tizio R. 2000. Factors affecting the development of stenospermic grape (Vitis vinifera) embryos cultured in vitroActa Horticulturae528, 667–671.

Puglisi D, Las Casas G, Ferlito F, Nicolosi E, Di Guardo M, Scollo F, Saitta G, La Malfa S, Gentile A, Distefano G. 2022. Parents’ selection affects embryo rescue, seed regeneration and the heredity of seedless trait in table grape breeding programs. Agriculture12, 1096.

Ramming D W, Emershad R L. 1982. In-ovulo embryo culture of seeded and seedless Vitis vinifera L. Hortscience17, 487.

Ramming D W, Emershad R L, Spiegel-Roy P, Sahar N, Baron I. 1990a. Embryo culture of early ripening seeded grape (Vitis vinifera) genotypes. Hortscience25, 339–342.

Ramming D W, Ledbetter C A, Tarailo R. 1990b. Hybridization of seedless grapes. VITISJournal of Grapevine Research29, 439–444.

Stout A B. 1936. Seedlessness in Grapes. New York State Agricultural Experiment Station Technology Bulletin, New York. pp. 1–68.

Tian L L, Wang Y J. 2008a. Seedless grape breeding for disease resistance by using embryo rescue. VITISJournal of Grapevine Research47, 15–19.

Tian L L, Wang Y J, Niu L, Tang D M. 2008b. Breeding of disease-resistant seedless grapes using Chinese wild Vitis spp. I. In vitro embryo rescue and plant development. Scientia Horticulturae117, 136–141.

Valdez J G. 2005. Immature embryo rescue of grapevine (Vitis vinifera L.) after an extended period of seed trace culture. VITISJournal of Grapevine Research44, 17–23.

Xu T F, Guo Y R, Wang W Y, Yuan X J, Chu Y N, Wang X W, Han Y L, Wang Y J, Song R, Fang Y L, Wang L J, Xu Y. 2022. Effects of exogenous paclobutrazol and sampling time on the efficiency of in vitro embryo rescue in the breeding of new seedless grape varieties. Journal of Integrative Agriculture, 21, 1633–1644.

Yamashita H, Haniuda T, Shiba H. 1995. In vitro culture of embryos obtained by crossing tetraploid grape cultivar ‘Kyoho’ with diploid cultivars. Journal of the Japanese Society for Horticultural Science63, 719–724.

Yamashita H, Shigehara I, Haniuda T. 1998. Production of triploid grapes by in ovulo embryo culture. VITISJournal of Grapevine Research37, 113–117.

Yang D L, Li W, Li S, Yang X L, Wu J L, Cao Z Y. 2007. In vitro embryo rescue culture of F1 progenies from crosses between diploid and tetraploid grape varieties. Plant Growth Regulation51, 63–71.

Zhu P P, Gu B, Li P Y, Shu X, Zhang X, Zhang J X. 2020. New cold-resistant, seedless grapes developed using embryo rescue and marker-assisted selection. Plant CellTissue and Organ Culture140, 551–562.

Zhu P P, Tian Y C, Liu Q Y, Ge Q Y, Zhang J X. 2022a. Optimisation of embryo rescue for cold-resistant seedless grapevine. New Zealand Journal of Crop and Horticultural Science52, 38–51.

Zhu P P, Zhang J X. 2022b. Effects of pre-bloom spraying thidiazuron and different embryo development media on seedless grape embryo rescue. New Zealand Journal of Crop and Horticultural Science53, 113–140.

[1] WANG Fu-qiang, BIAN Lu, QIU Peng-peng, GUO Shuo, GUO Jing-han, GUO Chen-shuo, JIANG Jian-fu, LIU Chong-huai, WANG Yong, LIU Guo-tian, WANG Yue-jin, XU Yan. Development and application of KASP marker for high throughput detection of the seedless trait in grapevine[J]. >Journal of Integrative Agriculture, 2023, 22(11): 3269-3283.
No Suggested Reading articles found!